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Growth Hormone Potentiates Colony Formation of Epiphyseal Chondrocytes in Suspension Culture^*

Department of Physiology, University of Göteborg S-400 33 Göteborg, Sweden

Address requests for reprints and all correspondence to: Dr. Anders Lindahl, Department of Physiology, University of Göteborg, P.O. Box 33031, S-400 33 Göteborg, Sweden.

Abstract

The effect of human GH (hGH) on colony formation of rat epiphyseal plate chondrocytes was studied in suspension culture. Chondrocytes were isolated enzymatically from epiphyseal plates of the proximal tibia of 28-day-old normal male rats, and were cultured in a suspension stabilized with 0.5% agarose. After approximately 7 days of culture in the presence of 10% newborn calf serum (NCS), chondrocyte colonies developed consisting of varying numbers of cells in matrix. No colonies developed in the absence of NCS, and the number of formed colonies was proportional to the concentration of NCS (5–20%) in the medium.

hGH potentiated the formation of large size colonies (diameter >90 µm) after a culture period of 10 or 14 days. The lowest effective concentration of hGH was 10 ng/ml, while 40 ng/ml hGH gave a maximal stimulatory effect (40–50%). Higher concentrations of hGH (80 and 160 ng/ml) showed reduced potentiation of colony formation. The stimulatory effect of hGH was expressed at 10–15% of NCS at 14 days of culture. There was a linear relation between the number of seeded cells and the number of colonies formed, both in the absence and presence of hGH.

These results show that GH potentiates colony formation in chondrocytes of the epiphyseal growth plate, providing further support for the contention that GH exert a direct stimulatory effect on epiphyseal cartilage and thus stimulates longitudinal bone growth directly. The finding that GH preferentially potentiated the formation of large size colonies suggests that GH promoted the differentiation of early proliferative chondrocytes or stem cell chondrocytes with an inherent high capacity to proliferate. (Endocrinology 118: 1843–1848, 1986)

Footnotes

^* This work was supported by grants from the Swedish Medical Research Council (14X-04250), The Göteborg Medical Society, and the Faculty of Medicine, University of Göteborg.

Received July 17, 1986.

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